A kind of nanoparticle and its preparation method and application

A nanoparticle and small interference technology, applied in the field of medicine, can solve the problems of high biological toxicity, difficulty in large-scale production, high production cost, etc., and achieve the effect of enhancing the inhibitory effect and promoting the proliferation and activation of T cells

Active Publication Date: 2021-02-12
INST OF PROCESS ENG CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

These carriers all have disadvantages such as high preparation cost, difficulty in large-scale production, or high biological toxicity.

Method used

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  • A kind of nanoparticle and its preparation method and application
  • A kind of nanoparticle and its preparation method and application
  • A kind of nanoparticle and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Embodiment 1: the preparation of nanoparticle vaccine

[0050] Preparation of LDH: quickly add 10mL magnesium nitrate (0.03M) and aluminum nitrate (0.01M) mixture into 40mL NaOH (0.15M) solution, stir rapidly at room temperature for 15min, then centrifuge at 12000g for 5min. After the obtained product was resuspended in decarbonated water, it was shaken and suspended at 30°C for 30 min, and the resulting suspension was transferred to a dialysis bag, and dialyzed in a closed container with decarbonated water 100 times the volume of the suspension at room temperature for 5 h. Then the suspension was transferred to an airtight container, shaken and suspended at 30°C for 1h at 280rpm, then transferred to a 100mL polytetrafluoroethylene hydrothermal kettle (50% sample volume), kept at 120°C for 15h, and cooled naturally.

[0051] Preparation of LDH / IDO-siRNA: Mix the prepared hydrotalcite with IDO siRNA, the sense strand (SEQ ID NO.1) of the IDO siRNA: 5'-GGGCUUCUUCCUCGUCUCU...

Embodiment 2

[0055] Example 2: Determination of the ability of IDO-siRNA to be inserted into LDH in vaccines

[0056] Agarose gel electrophoresis was performed according to conventional methods. In the gel electrophoresis diagram, free IDO siRNA migrated to the bottom of the gel, and IDO siRNA that entered the LDH layer remained in the sample well. The calculation method for the ability of IDO-siRNA intercalation into the LDH interlayer is as follows:

[0057] The IDO siRNA intercalation ability of hydrotalcite is calculated according to the following formula:

[0058]

[0059] Test results such as figure 1 As shown, the mass of free IDO siRNA in channel 1 is 1 μg, the mass of LDH in channel 2 is 40 μg, the mass of IDO siRNA in channels 3-8 is 1 μg, and the mass of LDH is added according to different mass ratios.

[0060]The results show that using the method described in the present invention to intercalate IDO siRNA to LDH, when the mass ratio of hydrotalcite and IDOsiRNA is 10:1, I...

Embodiment 3

[0061] Example 3: Determination of the ability of LDH / IDO-siRNA to adsorb Trp2

[0062] The conventional 2,2-biquinoline-4,4-dicarboxylate disodium assay method was used to determine the ability of LDH / IDO-siRNA to adsorb Trp2, and the adsorption capacity of LDH / IDO-siRNA to Trp2 was calculated according to the following formula:

[0063]

[0064] Description: Mg 3 The quality of Al-IDO siRNA-LDH is based on the mass of hydrotalcite

[0065] Test results such as figure 2 Shown:

[0066] When the mass of LDH / IDO-siRNA and Trp2 is 8:1, the adsorption capacity of Trp2 is 8.5%;

[0067] When the mass of LDH / IDO-siRNA and Trp2 is 8:2, the adsorption capacity of Trp2 is 13.9%;

[0068] When the mass of LDH / IDO-siRNA and Trp2 is 8:3, the adsorption capacity of Trp2 is 15.5%;

[0069] When the mass of LDH / IDO-siRNA and Trp2 is 8:4, the adsorption capacity of Trp2 is 17.8%;

[0070] When the mass of LDH / IDO-siRNA and Trp2 is 8:6, the adsorption capacity of Trp2 is 30.9%;

[...

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Abstract

The present invention relates to the field of medical technology, in particular to a nanoparticle and its preparation method and application. The nanoparticle uses a layered bishydroxy compound metal oxide as a carrier, and the carrier is loaded with small interfering RNA and an antigenic epitope peptide, wherein , the small interfering RNA is inserted into the carrier, and the antigen epitope peptide is adsorbed on the surface of the carrier. The nanoparticles of the present invention can co-deliver indoleamine 2,3-dioxygenase small interfering RNA and hydrotalcite nanoparticles of tyrosinase-related antigen peptide 2. The particles have the ability to target DC cells, break immune tolerance, activate The immune system, and the function of causing a strong cellular immune response, compared with the vaccine or IDO‑siRNA based on the tumor antigen-specific epitope Trp2 alone, the inhibitory effect on the growth of melanoma was significantly enhanced.

Description

technical field [0001] The invention relates to the technical field of medicine, in particular to a nanoparticle and its preparation method and application, in particular to the preparation and application of a hydrotalcite nanoparticle co-delivering small interfering RNA and tumor-associated epitope. Background technique [0002] Tyrosinase-related antigenic peptide 2 (Trp2) is a tumor-specific epitope, which can effectively inhibit the growth of melanoma by preparing it as a vaccine. Trp2 is a small molecular polypeptide composed of 9 amino acids, with a short sequence and weak immunogenicity, making it difficult to elicit effective toxic T cell responses. Therefore, carriers are usually used to enhance the immunogenicity of Trp2. Overexpression of IDO gene in dendritic cells can lead to an increase in the level of its downstream products IL-12 and TGF-β, and these cytokines are important factors that cause the body's immune tolerance. Indoleamine 2,3-dioxygenase small i...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K47/69A61K47/52A61K31/713A61K39/00A61K39/385A61P37/04A61P35/00
CPCA61K31/713A61K39/0011A61K39/385A61K2039/57A61K2039/62A61K2300/00
Inventor 刘瑞田张凌宵刘栋群
Owner INST OF PROCESS ENG CHINESE ACAD OF SCI
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